Application of specnuezhenide as autophagy inducer
A kind of special privet glycoside and inducer technology, applied in the field of medicine
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Embodiment 1
[0041] Example 1 The detection of teruzhenin in inducing autophagy of human lung cancer epithelial cells (A549)
[0042] Experimental materials: teruci, methicillin-resistant Staphylococcus aureus (MRSA), human lung cancer epithelial cells, that is, A549 cells (cultivated in DMEM medium containing 10% fetal bovine serum), DMEM was purchased from Hyclone Corporation of the United States , product number SH30022.01; fetal bovine serum was purchased from Shanghai Jitai Yikesai Biotechnology Co., Ltd., product number FND500; Ad-mCherry-GFP-LC3B adenovirus was purchased from Biyuntian Biotechnology Co., Ltd., product number C3011- 1ml), etc.
[0043] Establishment of autophagy flow detection model: first, in 96-well plate with 5×10 4 Inoculate A549 cells per well, 100uL per well, and culture for 24h. Ad-mCherry-GFP-LC3B adenovirus was used to infect A549 cells. The virus stock solution was diluted 60 times with serum-free medium to infect A549 cells at 25uL / well for 2 hours. At ...
Embodiment 2
[0057] Example 2 The detection of tercitrusin in inducing autophagy of human lung cancer epithelial cells (A549)
[0058] Establishment of autophagy flow detection model: first, in 96-well plate with 5×10 4 Inoculate A549 cells per well, 100uL per well, and culture for 24h. Ad-mCherry-GFP-LC3B adenovirus was used to infect A549 cells. The virus stock solution was diluted 60 times with serum-free medium to infect A549 cells at 25uL / well for 2 hours. At the same time, a group of A549 cells without Ad-mCherry-GFP-LC3B adenovirus was set as a blank control. In a 96-well plate, DMEM medium containing 3% serum was added to each well, 75uL / well. Continue culturing, adenovirus infection for 24 hours, replace the medium with fresh DMEM medium containing 3% serum, and continue culturing for 24 hours.
[0059] Establishment of MRSA-infected A549 cell model: In addition to the blank control group, the methicillin-resistant Staphylococcus aureus MRSA bacterial fluid was used for infecti...
Embodiment 3
[0062] Example 3 The detection of teruzhenin in inducing autophagy of human lung cancer epithelial cells (A549)
[0063] Establishment of autophagy flow detection model: first, in 96-well plate with 5×10 4 Inoculate A549 cells per well, 100uL per well, and culture for 24h. Ad-mCherry-GFP-LC3B adenovirus was used to infect A549 cells. The virus stock solution was diluted 60 times with serum-free medium to infect A549 cells at 25uL / well for 2 hours. At the same time, a group of A549 cells without Ad-mCherry-GFP-LC3B adenovirus was set as a blank control. In a 96-well plate, DMEM medium containing 3% serum was added to each well, 75uL / well. Continue culturing, adenovirus infection for 24 hours, replace the medium with fresh DMEM medium containing 3% serum, and continue culturing for 24 hours.
[0064] Establishment of MRSA-infected A549 cell model: In addition to the blank control group, the methicillin-resistant Staphylococcus aureus MRSA bacterial fluid was used for infectio...
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